中药材2011，Vol.34Issue(10)：1498-1502,5.
侵染浙贝母的浙贝母病毒Y(FVY)CP基因的原核表达及抗血清制备
Prokaryotic Expression of CP Gene of Fritillary virus Y Infecting Thunberg Fritillary and Antiserum Preparation
摘要
Abstract
Objective:To prepare antiserum against Fritillary virus Y(FVY) CP for detecting FVY and study serological relationships with other viruses.Methods:Specific primer was designed according to Genbank ( accession:AM039800) to amplify CP gene of FVY infecting Thunberg fritillary.Sequence relationship with other potyviruses was made by Blast.The CP gene was inserted into pSBET and expressed in Escherichia coli BL21 ( DE3 ) plys E strain.The object protein was purified by 12％ SDS-PAGE firstly and subsequently 5％-20％ gradient SDS-PAGE.The antiserum against the CP was raised in mouse and its specificity was confirmed by Western blot analysis.The reactivity of the antiserum produced to FVY CP was tested by Western blot against the over-expressed coat proteins of 17 potyviruses.The ability to combine with nature FVY particles was confirmed by ELISA analysis.Results:It shared 81.2％ nucleotide acids identities with TrVY(Tricyrtis virus Y,AY 864850) CP gene,68.1％ with SMV-P (Soybean mosaic virus Pinellia strain,AJ507388.2)CP gene and 67.2％ with ZYMV(Zucchini yellow mosaic virus Luan isolate) CP gene.The prepared antiserum was special to FVY CP,also reacted moderately to the expressed CP of SMV-P(Soybean mosaic virus Pinellia strain) and weakly to that of ZYMV (Zucchini yellow mosaic virus Luan isolate).Conclusion:The antibody could combine to nature FVY particles and the antiserum is suitable for FVY detection by ELISA in large scale.关键词
浙贝母/浙贝母病毒Y/CP基因/原核表达/抗血清制备Key words
Thunberg fritillary/Fritillary virus Y/CP gene/Prokaryotic expression/Antiserum preparation分类
生物科学引用本文复制引用
韦传宝,隗洋洋,杨宇,刘士亮,胡皓雨,何月..侵染浙贝母的浙贝母病毒Y(FVY)CP基因的原核表达及抗血清制备[J].中药材,2011,34(10):1498-1502,5.基金项目
安徽省教育厅自然科学基金重点项目资助(KJ2010A328 （）
J2011A272) （）
